Journal: Cell Communication and Signaling : CCS

Article Title: p53 regulates early mucosal immunity via antigen presentation in zebrafish intestine post-SVCV vaccination

doi: 10.1186/s12964-026-02749-8

Figure Lengend Snippet: P53 mediates the antigen presentation through TAP1 and ERAP1. A The expression of indictors involved in p53-mediated antigen presentation signaling pathways in intestine from vaccinated fish at 3 and 7 dpv. After treated with G3, intestine from vaccinated fish at 3 and 7 dpv were collected and the expression level of proteins was detected by western blotting. B Relative cell viability of intestine lymphocytes after incubation with different concentrations of 5-Fu for 48 h ( n = 5, each group with 3 replicates). C Relative cell viability of intestine lymphocytes after incubation with different concentrations of MG-132 for 48 h ( n = 5, each group with 3 replicates). D Relative cell viability of intestine lymphocytes after incubation with different concentrations of NH4Cl for 48 h ( n = 5, each group with 3 replicates). E Relative cell viability of intestine lymphocytes after incubation with different concentrations of PFT-μ for 48 h ( n = 5, each group with 3 replicates). F The expression of indictors involved in p53-mediated antigen presentation pathways when the proteasomal and lysosomal pathways were inhibited. After treated with G3 for 12 h, intestine lymphocytes were cultured with MG-132 or NH4Cl for 48 h. Then the cells were collected and the expression level of proteins was detected by western blotting. G The expression of indictors involved in p53-mediated antigen presentation pathways when p53 was activated and antigen presentation pathways were inhibited. After treated with G3 for 12 h, intestine lymphocytes were cultured with MG-132, NH4Cl, or 5-Fu for 48 h. Then the cells were collected and the expression level of proteins was detected by western blotting. H The expression of indictors involved in p53-mediated antigen presentation pathways when p53 was activated. After treated with G3 for 12 h, intestine lymphocytes were cultured with 5-Fu for 48 h. Then the cells were collected and the expression level of proteins was detected by western blotting. I The expression of indictors involved in p53-mediated antigen presentation pathways when p53 was inhibited. After treated with G3 for 12 h, intestine lymphocytes were cultured with PFT-μ for 48 h. Then the cells were collected and the expression level of proteins was detected by western blotting. J The knockdown effect of p53 was detected by qPCR ( n = 5, each group with 3 replicates). Zebrafish were injected with NC, p53-siRNA-1, p53-siRNA-2, or p53-siRNA-3. Then intestinal tissues were collected to isolate lymphocytes and the mRNA expression level of p53 was detected by qPCR. K The knockdown effect of p53 was detected by western blotting. Zebrafish were injected with NC, p53-siRNA-1, p53-siRNA-2, or p53-siRNA-3. Then intestinal tissues were collected to isolate lymphocytes and the protein expression level of p53 was detected by western blotting. L The expression of indictors involved in p53-mediated antigen presentation pathways when p53 was knock down. Zebrafish were immunized with G3 for 6 h, and then injected with p53-siRNA-3. The intestinal tissues were collected to isolate lymphocytes and the protein expression level was detected by western blotting. The P value for each study was determined by one-way ANOVA. * P < 0.05, ** P < 0.01. Data are representative of three independent experiments (means ± SEM)

Article Snippet: The mouse anti-β-actin (66009-1-Ig), mouse anti-LCK (32464-1-AP), rabbit anti-53 (10442-1-AP), rabbit anti-TAP1 (11114-1-AP), and rabbit anti-ERAP1 (13821-1-AP) antibodies were purchased from Proteintech (Wuhan, China).

Techniques: Immunopeptidomics, Expressing, Protein-Protein interactions, Western Blot, Incubation, Cell Culture, Knockdown, Injection